title: Purification and partial characterization of the major outer membrane protein of Neisseria gonorrhoeae.
abstract: A procedure is described to isolate the major outer membrane protein (protein I) from Neisseria gonorrhoeae in large quantities. The method involves precipitation of protein I by hexadecyltrimethylammonium bromide (CTB) at low ionic strength. CTB is lethal for the gonococci and solubilizes most other proteins. Protein I is brought into solution by raising the ionic strength, and the nucleic acids are subsequently removed by 20% ethanol precipitation. The CTB is removed by precipitating protein I with ethanol and replaced by N-tetradecyl-N,N-dimethyl-3-ammonia-1-propanesulfonate, a dipolar ionic detergent. Further purification is accomplished by ion-exchange and molecular sieve chromatography. Two species of protein I (34,000 daltons [34K] and 32K) were purified by these methods. The purified proteins reacted with antisera prepared against the homologous organisms. The 34K proteins I generated proteolytic fragments upon treatment with trypsin and chymotrypsin similar to those generated by 34K protein in intact gonococci. The amino acid compositions of the three proteins were much like those of other major proteins of gram-negative organisms.
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D008565:Membrane Proteins / Q000032:analysis / Q000302:isolation & purification*; D008970:Molecular Weight; D009344:Neisseria gonorrhoeae / Q000032:analysis*
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D002593:Cetrimonium Compounds; D008565:Membrane Proteins; D000077286:Cetrimonium